Abstract: The antioxidant activity and the chemical composition of methanol extracts from peel and pulp belonging to two species of Tunisian prickly pears Opuntia ficus indica (spiny and thornless forms) and Opuntia stricta has been studied. The antioxidant capacity was measured by DPPH radical scavenging activity. The Total Phenolic Compound (TPC) and the total flavonoid content were determined by the Folin-Ciocalteu method and colorimetric method, respectively. The phenolic compounds were identified and quantified by High Performance Liquid Chromatography (HPLC) coupled with an electrospray ionization mass spectrometry (ESI-MS). The results showed that O. stricta fruits present the best antioxidant activities than the two forms of O. ficus indica while the TPC was more important in O. ficus indica than in the O. stricta fruits. The peels have higher flavonoids than pulps and the thornless has more flavonoid than the spiny. The RP-HPLC and ESI-MS analysis detected two classes of phenolic compounds and betalain pigments. Isorhamnetin derivatives are the dominant flavonol glycoside identified in O. ficus indica (spiny: 65.25 μg g-1; thornless: 77.03 μg g-1) and O. stricta peels (19.22 μg g-1).